An optical path structure for detecting myocardial cell function with multimodal synchronous imaging

Abstract

The present invention relates to the technical field of multimodal synchronous imaging, and particularly relates to an optical path structure for detecting myocardial cell functions by multimodal synchronous imaging. The emission light excited by the sample to be photographed is filtered by a first fluorescence filter block, and the emission light is respectively filtered and reflected by a second fluorescence filter block, so as to form two beams of emission light at a first camera shooting port and a second camera shooting port. The incident light at the first camera shooting port and the second camera shooting port is collected by a first camera and a second camera respectively, so as to simultaneously complete the collection of mechanical data and calcium imaging of the sample to be photographed, thereby solving the problems existing in the prior art.

Description

Technical Field

[0001] The present invention relates to the technical field of multimodal synchronous imaging, and particularly to an optical path structure for detecting myocardial cell function by multimodal synchronous imaging. Background Art

[0002] The combination of calcium concentration imaging technology and nano-hydrogel mechanical analysis is a frontier research direction in the fields of biomedical engineering and materials science. Calcium imaging technology uses fluorescent probes or genetically encoded calcium indicators (such as GCaMP) to monitor the changes in intracellular calcium ion concentration in real time, and is widely used in fields such as neuroscience and cardiovascular disease research. Its core equipment relies on high-speed and high-sensitivity imaging systems, such as high-frame-rate cameras and large-field optical modules, to capture dynamic cell activities. Nano-hydrogels, due to their high water content, biocompatibility, and tunable mechanical properties, are used in fields such as drug delivery, tissue engineering, and biosensing. In recent years, researchers have attempted to use hydrogels as mechanical response carriers to analyze the mechanical properties of the microenvironment (such as hardness and viscoelasticity) through their deformation or conductivity changes.

[0003] However, in the prior art, calcium imaging devices and mechanical analysis systems usually operate independently, resulting in the following problems:

[0004] Data asynchronization: There is a time difference in the acquisition of calcium signals and mechanical signals, making it difficult to correlate dynamic biological processes with changes in the mechanical microenvironment; Equipment redundancy: Multiple sets of systems occupy a large space, are complex to operate, and increase experimental costs; Functional singularity: Calcium imaging devices (such as) focus on capturing optical signals and lack a mechanical sensing module, and cannot synchronously obtain material deformation or stress data. Mechanical analysis devices (such as atomic force microscopes) require contact measurement, which may interfere with cell activity or damage the hydrogel structure. Summary of the Invention

[0005] In view of this, the purpose of the present invention is to provide an optical path structure for detecting myocardial cell function by multimodal synchronous imaging to overcome the problems existing in the current prior art.

[0006] To achieve the above purpose, the present invention adopts the following technical solutions:

[0007] The present application provides an optical path structure for detecting myocardial cell function by multimodal synchronous imaging, including: a fluorescence light source, a microscope objective lens, a first fluorescence filter block, a reflector, a second fluorescence filter block, a first camera, a second camera, and a support structure;

[0008] The microscope objective lens is arranged at the sample focusing port of the support structure for focusing on and photographing a sample; wherein, the sample to be photographed is a cell sample spread on a hydrogel containing nano-fluorescent microspheres, and the cell sample has been subjected to fluorescence staining treatment;

[0009] The fluorescent light source is aligned with the light source inlet of the support structure to provide excitation light;

[0010] The first fluorescence filter block is disposed within the support structure at the light source inlet. The excitation light is filtered and reflected by the first fluorescence filter block, passes through the microscope objective lens, and irradiates the sample to be imaged;

[0011] The mirror is disposed at a corner of the support structure to reflect the emission light excited by the sample to be imaged after being irradiated by the excitation light; wherein, the emission light has been filtered by the first fluorescence filter block;

[0012] The support structure has a first camera shooting port and a second camera shooting port;

[0013] The second fluorescence filter block is disposed at the intersection of the first camera shooting port and the second camera shooting port to receive the emission light reflected by the mirror, filter and reflect the emission light, and cause the emission light to enter the first camera shooting port and the second camera shooting port respectively;

[0014] The first camera is aligned with the first camera shooting port to receive the emission light filtered by the second fluorescence filter block and complete the acquisition of mechanical data;

[0015] The second camera is aligned with the second camera shooting port to receive the emission light reflected and filtered by the second fluorescence filter block and complete calcium imaging.

[0016] Further, for the above optical path structure, the fluorescent light source is: a mercury lamp or a multi-color LED light source.

[0017] Further, for the above optical path structure, the first fluorescence filter block includes: a first filter block bracket, a DM505 filter, a BA510 - 550 filter, and a BP460 - 495 filter;

[0018] The DM505 filter, the BA510 - 550 filter, and the BP460 - 495 filter are disposed within the first filter block bracket, and the DM505 filter is disposed at a preset angle with respect to the BA510 - 550 filter and the BP460 - 495 filter respectively;

[0019] The excitation light is filtered by the BP460 - 495 filter, then reflected by the DM505 filter, passes through the microscope objective lens, and irradiates the sample to be imaged;

[0020] The emission light is filtered by the DM505 filter, and then filtered by the BA510 - 550 filter and irradiates the mirror.

[0021] Further, in the above-described optical path structure, the mirror is a total reflection mirror.

[0022] Further, in the above-described optical path structure, the second fluorescence filter block includes: a second filter block bracket, a BA510IF filter, a BA578 - 633 filter, and a BA510 - 550 filter;

[0023] The BA510IF filter, the BA578 - 633 filter, and the BA510 - 550 filter are disposed within the second filter block bracket, and the BA510IF filter is disposed at a preset angle with respect to the BA578 - 633 filter and the BA510 - 550 filter respectively;

[0024] The emitted light reflected by the mirror is filtered by the BA510IF filter and then filtered by the BA578 - 633 filter and shot towards the first camera shooting port;

[0025] The emitted light reflected by the mirror is also reflected by the BA510IF filter, filtered by the BA510 - 550 filter, and shot towards the second camera shooting port.

[0026] Further, in the above-described optical path structure, the first camera is a high quantum efficiency CMOS or SCMOS camera for photographing fluorescent microspheres.

[0027] Further, in the above-described optical path structure, the second camera is a high quantum efficiency CMOS or SCMOS camera for calcium imaging.

[0028] The beneficial effects of the present invention are:

[0029] This application includes a fluorescence light source, a microscope objective lens, a first fluorescence filter block, a reflector, a second fluorescence filter block, a first camera, a second camera, and a support structure; the microscope objective lens is disposed at the sample focusing port of the support structure for focusing on and photographing a sample; wherein, the sample to be photographed is a cell sample laid on a hydrogel containing nano-fluorescent microbeads, and the cell sample has been subjected to fluorescence staining treatment; the fluorescence light source is aligned with the light source inlet of the support structure for providing excitation light; the first fluorescence filter block is disposed within the support structure at the light source inlet, and the excitation light passes through the filtering and reflection of the first fluorescence filter block, passes through the microscope objective lens and irradiates the sample to be photographed; the reflector is disposed at the corner of the support structure for reflecting the emitted light excited by the sample to be photographed after being irradiated by the excitation light; wherein, the emitted light has passed through the filtering of the first fluorescence filter block; the support structure has a first camera photographing port and a second camera photographing port; the second fluorescence filter block is disposed at the intersection of the first camera photographing port and the second camera photographing port for receiving the emitted light reflected by the reflector, and filtering and reflecting the emitted light so that the emitted light is respectively incident into the first camera photographing port and the second camera photographing port; the first camera is aligned with the first camera photographing port for receiving the emitted light filtered by the second fluorescence filter block to complete the acquisition of mechanical data; the second camera is aligned with the second camera photographing port for receiving the emitted light reflected and filtered by the second fluorescence filter block to complete calcium imaging. In this application, the emitted light excited by the sample to be photographed is filtered by the first fluorescence filter block, and the emitted light is respectively filtered and reflected by the second fluorescence filter block, so as to form two emitted lights at the first camera photographing port and the second camera photographing port, and the incident light at the first camera photographing port and the second camera photographing port is respectively collected by the first camera and the second camera, so as to simultaneously complete the acquisition of the mechanical data and calcium imaging of the sample to be photographed, thereby solving the problems existing in the prior art. Description of the Drawings

[0030] In order to more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the following will briefly introduce the drawings required for use in the description of the embodiments or the prior art. Obviously, the following drawings are only some embodiments of the present invention, and for those of ordinary skill in the art, other drawings can be obtained based on these drawings without creative efforts.

[0031] Figure 1 It is a schematic structural diagram provided by an embodiment of an optical path structure for detecting myocardial cell functions with multimodal synchronous imaging of the present invention;

[0032] Figure 2 It is a schematic structural diagram of a first fluorescence filter block provided by an embodiment of an optical path structure for detecting myocardial cell functions with multimodal synchronous imaging of the present invention;

[0033] Figure 3It is a schematic diagram of the second fluorescence filter block structure provided by an embodiment of an optical path structure for detecting myocardial cell function with multimodal synchronous imaging according to the present invention. Detailed implementation manners

[0034] To make the objectives, technical solutions and advantages of the present invention clearer, the technical solutions of the present invention will be described in detail below. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all the embodiments. All other implementation manners obtained by those of ordinary skill in the art based on the embodiments of the present invention without creative efforts shall fall within the scope protected by the present invention.

[0035] As Figure 1 shown, Figure 1 It is a schematic diagram of a structure provided by an embodiment of an optical path structure for detecting myocardial cell function with multimodal synchronous imaging according to the present invention. This embodiment may include:

[0036] A fluorescence light source 1, a microscope objective lens 2, a first fluorescence filter block 4, a mirror 9, a second fluorescence filter block 6, a first camera 7, a second camera 8, and a support structure 5;

[0037] The microscope objective lens 2 is arranged at the sample focusing port of the support structure 5 and is used for focusing on and photographing the sample 3; wherein, the photographed sample 3 is a cell sample laid on a hydrogel containing nano-fluorescent microbeads, and the cell sample has been subjected to fluorescence staining treatment;

[0038] The fluorescence light source 1 is aligned with the light source inlet of the support structure 5 and is used for providing excitation light 10;

[0039] The first fluorescence filter block 4 is arranged inside the support structure 5 and at the light source inlet. The excitation light 10 passes through the filtration and reflection of the first fluorescence filter block 4, passes through the microscope objective lens 2, and irradiates the photographed sample 3;

[0040] The mirror 9 is arranged at the corner of the support structure 5 and is used for reflecting the emission light 11 excited by the photographed sample 3 after being irradiated by the excitation light 10; wherein, the emission light 11 has passed through the filtration of the first fluorescence filter block 4;

[0041] The support structure 5 has a photographing port for the first camera 7 and a photographing port for the second camera 8;

[0042] The second fluorescence filter block 6 is arranged at the intersection of the photographing port for the first camera 7 and the photographing port for the second camera 8, and is used for receiving the emission light 11 reflected by the mirror 9, filtering and reflecting the emission light 11, and making the emission light 11 enter the photographing port for the first camera 7 and the photographing port for the second camera 8 respectively;

[0043] The first camera 7 is aligned with the photographing port for the first camera 7 and is used for receiving the emission light 11 filtered by the second fluorescence filter block 6 to complete the acquisition of mechanical data;

[0044] The second camera 8 is aligned with the second camera 8 shooting port for receiving the emitted light 11 that has been reflected and filtered by the second fluorescence filter block 6, completing calcium imaging.

[0045] It can be understood that this embodiment has a fluorescence light source 1, a microscope objective lens 2, a first fluorescence filter block 4, a mirror 9, a second fluorescence filter block 6, a first camera 7, a second camera 8, and a support structure 5; the microscope objective lens 2 is arranged at the sample focusing port of the support structure 5 for focusing on and shooting the sample 3; wherein, the sample 3 to be shot is a cell sample laid on a hydrogel containing nano-fluorescent microbeads, and the cell sample has been subjected to fluorescence staining treatment; the fluorescence light source 1 is aligned with the light source inlet of the support structure 5 for providing the excitation light 10; the first fluorescence filter block 4 is arranged inside the support structure 5 at the light source inlet, and the excitation light 10 passes through the filtering and reflection of the first fluorescence filter block 4, passes through the microscope objective lens 2 and shoots towards the sample 3 to be shot; the mirror 9 is arranged at the corner of the support structure 5 for reflecting the emitted light 11 excited by the sample 3 after being irradiated by the excitation light 10; wherein, the emitted light 11 has passed through the filtering of the first fluorescence filter block 4; the support structure 5 has a first camera 7 shooting port and a second camera 8 shooting port; the second fluorescence filter block 6 is arranged at the intersection of the first camera 7 shooting port and the second camera 8 shooting port for receiving the emitted light 11 reflected by the mirror 9 and filtering and reflecting the emitted light 11, so that the emitted light 11 is respectively incident into the first camera 7 shooting port and the second camera 8 shooting port; the first camera 7 is aligned with the first camera 7 shooting port for receiving the emitted light 11 that has been filtered by the second fluorescence filter block 6, completing the acquisition of mechanical data; the second camera 8 is aligned with the second camera 8 shooting port for receiving the emitted light 11 that has been reflected and filtered by the second fluorescence filter block 6, completing calcium imaging. In this embodiment, the emitted light 11 excited by the sample 3 to be shot is filtered by the first fluorescence filter block 4, and the emitted light 11 is respectively filtered and reflected by the second fluorescence filter block 6, so as to form two emitted lights 11 at the first camera 7 shooting port and the second camera 8 shooting port, and the first camera 7 and the second camera 8 respectively collect the incident light at the first camera 7 shooting port and the second camera 8 shooting port, so as to simultaneously complete the acquisition of the mechanical data and calcium imaging of the sample 3 to be shot, thus solving the problems existing in the prior art.

[0046] Preferably, the fluorescence light source 1 is: a mercury lamp or a multi-color LED light source.

[0047] Preferably, the first fluorescence filter block 4 includes: a first filter block bracket, a DM505 filter, a BA510 - 550 filter, and a BP460 - 495 filter;

[0048] The DM505 filter, BA510-550 filter, and BP460-495 filter are arranged inside the first filter block bracket, and the DM505 filter is arranged at a preset angle with the BA510-550 filter and the BP460-495 filter respectively;

[0049] The excitation light 10 is filtered by the BP460-495 filter, and then reflected by the DM505 filter and passes through the microscope objective lens 2 and shoots towards the sample 3 to be photographed;

[0050] The emitted light 11 is filtered by the DM505 filter, and then filtered by the BA510-550 filter and shoots towards the mirror 9.

[0051] It can be understood that Figure 2 is a schematic diagram of the structure of the first fluorescence filter block 4 provided by an embodiment of an optical path structure for detecting myocardial cell function in multimodal synchronous imaging of the present invention. As Figure 2 shown, 401 is the first filter block bracket, 402 is the DM505 filter, 403 is the BA510-550 filter, and 404 is the BP460-495 filter.

[0052] The DM505 filter 402 is used to reflect light with a wavelength lower than 505 nm and transmit light with a wavelength greater than 505 nm. However, it is not limited to this model as long as the requirements can be met;

[0053] The BA510-550 filter 403 is used to transmit light in the range of 510-550 nm and block light with other wavelengths. However, it is not limited to this model as long as the requirements can be met;

[0054] The BP460-495 filter 404 is used to transmit light in the range of 460-495 nm and block light with other wavelengths. However, it is not limited to this model as long as the requirements can be met.

[0055] The excitation light 10 emitted by the fluorescent light source 1 can be light of a wide spectrum or narrow band. After being filtered by the BP460-495 filter 404 and then reflected by the DM505 filter 402, it further passes through the microscope objective lens 2, and the further light shoots towards the sample 3 to be photographed.

[0056] Preferably, the mirror 9 is a total reflection mirror.

[0057] Preferably, the second fluorescence filter block 6 includes: a second filter block bracket, a BA510IF filter, a BA578-633 filter, and a BA510-550 filter;

[0058] The BA510IF filter, BA578 - 633 filter, and BA510 - 550 filter are arranged within the second filter block bracket. The BA510IF filter is arranged at a preset angle with respect to the BA578 - 633 filter and the BA510 - 550 filter respectively;

[0059] The emitted light 11 reflected by the mirror 9, after being filtered by the BA510IF filter, is then filtered by the BA578 - 633 filter and directed towards the shooting port of the first camera 7;

[0060] The emitted light 11 reflected by the mirror 9 is also reflected by the BA510IF filter and filtered by the BA510 - 550 filter and then directed towards the shooting port of the second camera 8.

[0061] It can be understood that Figure 3 is a schematic diagram of the structure of the second fluorescence filter block 6 provided by an embodiment of the optical path structure for detecting myocardial cell function in multimodal synchronous imaging of the present invention. As Figure 3 shown, 601 is the second filter block bracket, 602 is the BA510IF filter, 603 is the BA578 - 633 filter, and 604 is the BA510 - 550 filter.

[0062] Preferably, the first camera 7 is: a high quantum efficiency CMOS or SCMOS camera, used for shooting fluorescent microspheres.

[0063] It can be understood that the emitted light 11 excited by the excitation of the sample 3 passes through the microscope objective lens 2, DM505 filter 402, BA510 - 550 filter 403, mirror 9, BA510IF filter 602, BA578 - 633 filter 603 in sequence. The further optical signal is finally received by the first camera 7 to complete the acquisition of mechanical data.

[0064] Preferably, the second camera 8 is: a high quantum efficiency CMOS or SCMOS camera, used for shooting calcium imaging.

[0065] It can be understood that the emitted light 11 excited by the excitation of the sample 3 passes through the microscope objective lens 2, DM505 filter 402, BA510 - 550 filter 403, mirror 9 in sequence, is further reflected by the BA510IF filter 602, and further passes through the BA510 - 550 filter 604. The optical signal is finally received by the second camera 8 to complete calcium imaging.

[0066] It can be understood that the same or similar parts in the above - mentioned embodiments can be referred to each other. For the content not detailed in some embodiments, reference can be made to the same or similar content in other embodiments.

[0067] It should be noted that in the description of the present invention, terms such as "first" and "second" are only used for descriptive purposes and cannot be construed as indicating or implying relative importance. In addition, in the description of the present invention, unless otherwise specified, the meaning of "a plurality of" refers to at least two.

[0068] Any process or method description in a flowchart or described otherwise herein can be understood to represent a module, segment, or portion of code including one or more executable instructions for implementing a specific logical function or process. The scope of the preferred embodiments of the present invention includes additional implementations where functions may be executed in a substantially simultaneous manner or in an order opposite to that shown or discussed, according to the functions involved, which should be understood by those skilled in the art to which the embodiments of the present invention pertain.

[0069] It should be understood that various parts of the present invention can be implemented by hardware, software, firmware, or a combination thereof. In the above embodiments, multiple steps or methods can be implemented by software or firmware stored in a memory and executed by a suitable instruction execution system. For example, if implemented by hardware, as in another embodiment, any one or a combination of the following techniques well known in the art can be used: discrete logic circuits having logic gate circuits for implementing logical functions on data signals, application specific integrated circuits having appropriate combinational logic gate circuits, programmable gate arrays (PGAs), field programmable gate arrays (FPGAs), etc.

[0070] Those of ordinary skill in the art of the present technology can understand that all or part of the steps carried by the methods of the above embodiments can be completed by instructing relevant hardware through a program. The said program can be stored in a computer-readable storage medium. When the program is executed, it includes one or a combination of the steps of the method embodiments.

[0071] In addition, each functional unit in various embodiments of the present invention can be integrated into a processing module, or each unit can exist physically alone, or two or more units can be integrated into one module. The above integrated module can be implemented in the form of hardware or in the form of a software functional module. When the integrated module is implemented in the form of a software functional module and sold or used as an independent product, it can also be stored in a computer-readable storage medium.

[0072] The above-mentioned storage medium can be a read-only memory, a magnetic disk, an optical disc, etc.

[0073] In the description of this specification, the descriptions referring to terms such as "one embodiment", "some embodiments", "examples", "specific examples", or "some examples", etc. mean that the specific features, structures, materials, or characteristics described in connection with the embodiment or example are included in at least one embodiment or example of the present invention. In this specification, the schematic descriptions of the above terms do not necessarily refer to the same embodiment or example. Moreover, the specific features, structures, materials, or characteristics described can be combined in a suitable manner in any one or more embodiments or examples.

[0074] Although the embodiments of the present invention have been shown and described above, it can be understood that the above embodiments are exemplary and should not be construed as limiting the present invention. Those of ordinary skill in the art can make changes, modifications, substitutions, and variations to the above embodiments within the scope of the present invention.

Claims

1. An optical path structure for detecting myocardial cell function with multimodal synchronous imaging, characterized in that, Comprising: A fluorescent light source, a microscope objective lens, a first fluorescence filter block, a reflector, a second fluorescence filter block, a first camera, a second camera, and a support structure; The microscope objective lens is disposed at the sample focusing port of the support structure for focusing on and photographing a sample; wherein, the sample to be photographed is a cell sample laid on a hydrogel containing nano-fluorescent microbeads, and the cell sample has been subjected to fluorescence staining treatment; The fluorescent light source is aligned with the light source inlet of the support structure for providing excitation light; The first fluorescence filter block is disposed within the support structure at the light source inlet, and the excitation light is filtered and reflected by the first fluorescence filter block and passes through the microscope objective lens and irradiates the sample to be photographed; The reflector is disposed at a corner of the support structure for reflecting the emission light excited by the sample to be photographed after being irradiated by the excitation light; wherein, the emission light has been filtered by the first fluorescence filter block; The support structure has a first camera photographing port and a second camera photographing port; The second fluorescence filter block is disposed at the intersection of the first camera photographing port and the second camera photographing port for receiving the emission light reflected by the reflector, and filtering and reflecting the emission light so that the emission light is respectively incident into the first camera photographing port and the second camera photographing port; The first camera is aligned with the first camera photographing port for receiving the emission light filtered by the second fluorescence filter block to complete the acquisition of mechanical data; The second camera is aligned with the second camera photographing port for receiving the emission light reflected and filtered by the second fluorescence filter block to complete calcium imaging.

2. The optical path structure according to claim 1, wherein The fluorescent light source is: a mercury lamp or a multi-color LED light source.

3. The optical path structure according to claim 2, wherein The first fluorescence filter block includes: a first filter block bracket, a DM505 filter, a BA510 - 550 filter, and a BP460 - 495 filter; The DM505 filter, the BA510 - 550 filter, and the BP460 - 495 filter are disposed within the first filter block bracket, and the DM505 filter is disposed at a preset angle with respect to the BA510 - 550 filter and the BP460 - 495 filter respectively; The excitation light is filtered by the BP460 - 495 filter and then reflected by the DM505 filter and passes through the microscope objective lens and irradiates the sample to be photographed; The emission light is filtered by the DM505 filter and then filtered by the BA510 - 550 filter and irradiates the reflector.

4. The optical path structure according to claim 3, wherein, The reflector is a total reflection reflector.

5. The optical path structure according to claim 4, characterized in that, The second fluorescence filter block includes: a second filter block bracket, a BA510IF filter, a BA578 - 633 filter, and a BA510 - 550 filter; The BA510IF filter, the BA578 - 633 filter, and the BA510 - 550 filter are disposed within the second filter block bracket, and the BA510IF filter is disposed at a preset angle with respect to the BA578 - 633 filter and the BA510 - 550 filter respectively; The emitted light reflected by the mirror is filtered by the BA510IF filter and then filtered by the BA578-633 filter and directed towards the first camera shooting port; The emitted light reflected by the mirror is also reflected by the BA510IF filter and filtered by the BA510-550 filter and directed towards the second camera shooting port.

6. The optical path structure according to claim 5, wherein The first camera is a high quantum efficiency CMOS or SCMOS camera for shooting fluorescent microspheres.

7. The optical path structure according to claim 6, characterized in that, The second camera is a high quantum efficiency CMOS or SCMOS camera for calcium imaging shooting.

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